KR0163671B1 - N-arylsulfonyl-alpha-propargylglycinamide derivatives - Google Patents

Abstract

The present invention comprises a novel N-arylsulfonyl-α-propargylglycinamide derivative of the general formula (I) shown below that is easy to synthesize and exhibits excellent thrombin inhibitory activity, a preparation method thereof, and a compound (I) as an active ingredient. It relates to a composition for preventing blood clotting or treating various thrombosis.

Wherein R ¹ is in each occurrence a substituted or unsubstituted aryl or heteroaryl, R ² and R ³ may be the same or different from each other, and each independently hydrogen, lower alkyl, cycloalkyl, or in each case Aryl or heteroaryl substituted or unsubstituted, R ² and R ³ are bonded together with the nitrogen atom to which they are attached to be substituted or unsubstituted by lower alkyl, carboxyl or carboalkoxy, and Containing 4 to 8-membered rings, R 4 represents lower alkyl, lower aminoalkyl, or in each case substituted or unsubstituted aryl, arylalkyl, heteroaryl or heteroarylalkyl.

Description

N-arylsulfonyl-α-propargylglycineamide derivatives

The present invention relates to nonpeptidic compounds useful as thrombin inhibitors.

More specifically, the present invention provides a novel N-arylsulfonyl-α-propargylglycinamide derivative of the general formula (I) below, which is easy to synthesize and exhibits excellent thrombin inhibitory activity, a preparation method thereof, and a compound (I) It relates to a composition for preventing blood clotting or treating various thrombosis containing as a component.

Wherein R 1 is in each case a substituted or unsubstituted aryl or heteroaryl, R 2 and R 3 may be the same or different from each other, and each independently hydrogen, lower alkyl, cycloalkyl, or in each case 4 represents unsubstituted aryl or heteroaryl, or R ² and R ³ are bonded together with the nitrogen atom to which they are attached to be substituted or unsubstituted by lower alkyl, carboxyl or carboalkoxy and contain 4 nitrogen atoms. To 8-membered ring, R ⁴ represents lower alkyl, lower aminoalkyl, or in each case substituted or unsubstituted aryl, arylalkyl, heteroaryl or heteroarylalkyl.

Several complex enzymatic reactions are involved in the coagulation mechanism, and the final stage involves the conversion of prothrombin to thrombin. Thrombin produced in this reaction plays an important role in blood coagulation, activating platelets and converting fibrin to fibrin, which is converted into a polymer in the polymerization reaction and crosslinked by activated blood factor XIII to insoluble thrombi. Becomes Thrombin also activates blood factors V and shock, which accelerate the blood coagulation reaction by feedback. Thus, inhibitors of thrombin may act as effective anticoagulants by inhibiting the activation of platelets and preventing the production of fibrin.

Peptides with cleavage sites similar to fibrin, the substrate of thrombin, are three amino acids, D-Phe-Pro-Arg, and derivatives that mimic this have been reported to be effective as thrombin inhibitors. These derivatives are compounds in which the C-terminal portion of D-Phe-Pro-Arg is substituted with aldehyde, phosphonate or boronic acid (Trends in Pharm. Sci. 1993, 14, 366-376). However, most of these compounds retain the properties of peptides, so they have good thrombin inhibitory effects but are not suitable for oral administration. These peptide compounds are generally unstable in vivo, poorly absorbed into organs, and degrade quickly. This is because it has a property. Therefore, it is very difficult to develop a peptide-based compound as a useful thrombin agent.

Therefore, in recent years, many non-peptidic compounds have been developed as thrombin inhibitors. As a representative non-peptidic thrombin inhibitor, agatroban (former US Pat. Nos. 94258192 and 4201863) of the following structural formula (A) commercialized in Japan was developed in 1990.

Agatroban (A), however, is an arylsulfonyl arginine-based compound, which has excellent efficacy as an injection, but has no activity for oral administration, and has a disadvantage of being obtained through a complicated synthesis process.

Accordingly, the present inventors have conducted intensive studies to develop a non-peptide thrombin inhibitor compound having a novel structure that can be easily synthesized and exhibit effective thrombin inhibitory activity, and as a result, the compound of general formula (I) as defined above It has been confirmed that this object is achieved and the present invention has been completed.

It is therefore an object of the present invention to provide a thrombin inhibitor of the novel structure of formula (I), which has the potential of oral administration.

It is still another object of the present invention to provide a method for preparing the compound of formula (I).

A third object of the present invention is to provide a pharmaceutical composition for preventing blood clotting or treating various thrombosis, containing the compound of the general formula (I) as an active ingredient.

According to the present invention there is provided novel N-arylsulfonyl-α-propargylglycineamide derivatives represented by the following general formula (I), pharmaceutically acceptable salts, hydrates, solvates and isomers thereof.

Wherein R ¹ is in each occurrence a substituted or unsubstituted aryl or heteroaryl, R ² and R ³ may be the same or different from each other, and each independently hydrogen, lower alkyl, cycloalkyl, or in each case Aryl or heteroaryl substituted or unsubstituted, R ² and R ³ are bonded together with the nitrogen atom to which they are attached to be substituted or unsubstituted by lower alkyl, carboxyl or carboalkoxy, and Containing 4 to 8-membered rings, R ⁴ represents lower alkyl, lower aminoalkyl, or in each case substituted or unsubstituted aryl, arylalkyl, heteroaryl or heteroarylalkyl.

As used herein to define a substituent of a compound of formula (I), the term loweralkyl used means straight or branched chain alkyl of 1 to 4 carbon atoms including methyl, ethyl, isopropyl, isobutyl, t-butyl do.

The term cycloalkyl means cyclic alkyl of 4 to 8 carbon atoms including cyclopentyl.

The term aryl means a 6-membered monocyclic aromatic group or a 10-membered bicyclic aromatic group, which groups are lower alkyl, amino, alkylamino, amidine, cyano, hydroxy, carboalkoxy, lower alkoxy Or optionally substituted with halogen.

The term heteroaryl also refers to a 5-6 membered monocyclic aromatic group or a 9 membered bicyclic aromatic group containing 1 to 3 heteroatoms selected from oxygen, nitrogen and sulfur, which groups are lower alkyl Optionally substituted by, amino, alkylamino, amidine, cyano, hydroxy, carboalkoxy, lower alkoxy or halogen.

Among the compounds of the formula (I) according to the invention, preferred compounds are those in which R ¹ represents aryl, R ² and R ³ each independently represent hydrogen, lower alkyl or cycloalkyl, or R ² and R ³ to which they are attached To form a 6-membered ring together with the nitrogen atom, and R ⁴ represents a lower alkyl or aryl or heteroaryl unsubstituted or substituted by an amino group.

Particularly preferred among the compounds of the general formula (I) according to the invention are those in which R ¹ represents naphthyl and R ² and R ³ each independently represent hydrogen, methyl or cyclopentyl, or R ² and R ³ are A combination with the attached nitrogen atom to form a piperidine group, R ⁴ is a compound representing phenyl, pyridyl, pyrimidinyl, pyrazolyl or imidazolyl unsubstituted or substituted by methyl or an amino group .

According to the invention representative compounds in the compounds of the general formula (I) include the following substances:

1) (S) -2- (naphthalene-2-sulfonylamino) -5-phenyl-pent-4-inoic acid dimethylamide

2) (S) -2- (naphthalene-2-sulfonylamino) -5-pyridin-4-yl-pent-4-inoic acid dimethylamide

3) (S) -2- (naphthalene-2-sulfonylamino) -5-pyridin-3-yl-pent-4-inoic acid dimethylamide

4) (S) -2- (naphthalene-2-sulfonylamino) -5-pyrimidin-5-yl-pent-4-inoic acid dimethylamide

5) 5- (6-Amino-pyridin-3-yl)-(S) -2- (naphthalene-2-sulfonylamino) -pent-4-inoic acid dimethylamide

6) 5- (2-Amino-pyrimidin-5-yl)-(S) -2- (naphthalene-2-sulfonylamino) -pent-4-inoic acid dimethylamide

7) (S) -2- (naphthalene-2-sulfonylamino) -5- (1H-pyrazol-4-yl) -pent-4-inoic acid dimethylamide

8) 5- (1H-imidazol-4-yl)-(S) -2- (naphthalene-2-sulfonylamino) -pent-4-inoic acid dimethylamide hydrochloride

9) (S) -2- (naphthalene-2-sulfonylamino) -5-phenyl-pent-4-inoic acid cyclopentyl-methyl-amide

10) (S) -2- (naphthalene-2-sulfonylamino) -5-pyridin-3-yl-pent-4-inoic acid cyclopentyl-methyl-amide

11) 5- (6-Amino-pyridin-3-yl)-(S) -2- (naphthalene-2-sulfonylamino) -pent-4-inoic acid cyclopentyl-methyl-amide

12) 5- (3-Amino-phenyl)-(S) -2- (naphthalene-2-sulfonylamino) -pent-4-inoic acid cyclopentyl-methyl-amide hydrochloride

13) 5- (4-Amino-phenyl)-(S) -2- (naphthalene-2-sulfonylamino) -pent-4-inoic acid cyclopentyl-methyl-amide trifluoroacetic acid salt

14) 5- (1H-imidazol-4-yl)-(S) -2- (naphthalene-2-sulfonylamino) -pent-4-inoic acid cyclopentyl-methyl-amide hydrochloride

15) Naphthalene-2-sulfonic acid [4- (3-amino-phenyl)-(S) -1- (piperidine-1-carbonyl) -but-3-ynyl] -amide hydrochloride

16) Naphthalene-2-sulfonic acid [4- (1H-amino-4-yl)-(S) -1- (piperidine-1-carbonyl) -but-3-ynyl] -amide hydrochloride

17) (S) -2- (naphthalene-2-sulfonylamino) -hex-4-inoic acid cyclopentyl-methyl-amide

18) 7-Amino- (S) -2- (naphthalene-2-sulfonylamino) -hept-4-inoic acid cyclopentyl-methyl-amide

The compounds of formula (I) according to the invention may also form pharmaceutically acceptable salts. Such pharmaceutically acceptable salts include acids that form non-toxic acid addition salts containing pharmaceutically acceptable anions such as inorganic acids, such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrobromic acid, hydroiodic acid, tartaric acid, formic acid, Organic carbonic acid such as citric acid, acetic acid, trichloroacetic acid or trifluoroacetic acid, gluconic acid, benzoic acid, lactic acid, fumaric acid, maleic acid, etc., sulfonic acid such as methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid or naphthalenesulfonic acid Acid addition salts formed by and the like are included.

The compounds of the present invention may have an asymmetric carbon center and may exist as racemates, racemic compounds, diastereomeric mixtures and individual diastereomers, all of these isomeric forms being included in the present invention.

The present invention also relates to a process for preparing the compound of the general formula (I). According to the present invention, the target compound of general formula (I) can be prepared by a method characterized by reacting an acetylene compound of general formula (II) with a compound of general formula (III). The production method of the present invention can be represented by the following scheme 1.

Scheme

Wherein R ¹ , R ² , R ³ and R ⁴ are each as defined above and X represents halogen.

Scheme 1 shows a process for preparing the final target compound from the intermediate acetylene compound (II) by one step process.

According to the process of the present invention, first, when R ⁴ is an aromatic group such as substituted or unsubstituted aryl, arylalkyl, heteroaryl or heteroarylalkyl, the intermediate acetylene compound (II) is cooled to room temperature in the presence of copper iodide and palladium catalyst. At the elevated temperature, the final target compound (I) is obtained by reacting with a haloaryl compound corresponding to R ⁴ to be introduced, preferably at a temperature of 25 ° C to 67 ° C. Palladium catalysts that may be used include bis (triphenylphosphine) palladium (II) chloride, tetrakis (triphenylphosphine) palladium (0), palladium (II) acetate, and the like, but are not limited thereto. no.

On the other hand, when R ⁴ is alkyl or lower aminoalkyl, the intermediate acetylene compound (II) is first treated with a strong base and then reacted with a haloalkane compound corresponding to R ⁴ under cooling, preferably at a temperature of -78 ° C to 0 ° C. To obtain the desired compound of general formula (I). Bases that can be used in this reaction include, but are not limited to, lithium hexamethyldisilazide (LHMDS), sodium hydride (NaH), butyllithium, and the like.

After completion of the reaction, the product can be separated and purified by conventional post-treatment methods such as chromatography, recrystallization and the like. In addition, the resultant compound of general formula (I) can be converted into an acid addition salt by treating with a suitable acid as mentioned above by conventional methods, if necessary.

The acetylene compounds of formula (II) used as intermediates in the preparation of compounds of formula (I) of the present invention in Scheme 1 are novel compounds and are therefore included within the scope of the present invention. This intermediate acetylene compound (II) can be prepared by the method shown in Scheme 2 below.

Scheme 1

Wherein R ¹ , R ² and R ³ are as defined above, and Boc means a t-butyloxycarbonyl group.

As shown in Scheme 2, the N-terminus of propargylglycine is first protected with a t-butyloxycarbonyl group (Boc) and the C-terminus is subjected to peptide coupling with the desired amine, followed by Treatment with trifluoroacetic acid removes the Boc-protecting group and the N-terminus of the resulting compound (4) can be coupled with arylsulfonylchloride (5) to produce the desired intermediate compound (II).

Coupling reagents known in the art that can be used for peptide coupling the C-terminus of Compound (2) with an amine compound in the above reaction include dicyclohexylcarbodiimide (DCC), 3-ethyl-3'- (Dimethylamino) -propylcarbodiimide (EDC), bis- (2-oxo-3-oxazolidinyl) -phosphinic chloride (BOP-C1), diphenylphosphoryl azide (DPPA), and the like, It is not limited to these. In addition, although the carboxylic acid compound (2) may be used as it is for this coupling reaction, it is preferable to further promote the reaction by converting it into an activating ester derivative to use in the coupling reaction. Activated ester derivatives of the carboxylic acid (2) are necessary to form amide bonds by coupling reactions with amines or to form ester bonds by coupling reactions with alcohols. Such activated ester derivatives are prepared by activating a carboxyl group by methods conventional in the art, specific examples of which include coupling with alkoxycarbonyl halides including methoxycarbonyl chloride, isobutyloxycarbonyl chloride, and the like. Carboxylic anhydrides derived from reagents, N-hydroxyphthalimide-derived esters, N-hydroxysuccinimide-derived esters, N-hydroxy-5-norbornene-2 ', 3'-di Conventional activated ester derivatives, such as carboxyimide-derived esters, 2,4,5-trichlorophenol-derived esters, are included, but are not limited to these.

As mentioned above, the compound of the general formula (I) according to the present invention is a compound exhibiting an excellent thrombin inhibitory effect. Therefore, the compounds of the present invention are useful for preventing blood clotting and for treating various thrombosis.

Therefore, another object of the present invention is to provide a pharmaceutical composition for preventing blood coagulation and for treating various thrombosis, containing a compound of formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient.

The total daily dose to be given to the host in a single dose or in separate doses when administering a compound of the present invention for clinical purposes is from 0.001 mg to 10 mg per kg of body weight in an adult, and the specific dose level for a particular patient is the specific compound to be used. May vary depending on body weight, sex, health status, diet, time of administration, method of administration, excretion rate, drug mixing and severity of disease.

The compounds of the present invention can be administered in injectable and oral formulations as desired. Injectable preparations, for example sterile injectable aqueous or oleaginous suspensions, can be prepared using suitable dispersing agents, wetting agents or suspending agents according to known techniques. Aqueous solvents that can be used include water, Ringer's solution and isotonic NaCl solution, and sterile fixed oils are commonly used as solvents or suspending media. Any non-irritating fixed oil may be used for this purpose, including mono- and diglycerides, and fatty acids such as oleic acid are used in the preparation of injectables.

Solid dosage forms for oral administration may be capsules, tablets, pills, powders, and granules. Particularly, capsules and tablets may be useful, and tablets and pills may be useful as enteric preparations. Solid dosage forms can be prepared by mixing the active compound with one or more inert diluents, such as sucrose, lactose, starch, and the like, lubricants such as magnesium stearate, disintegrants, binders and the like.

The compounds according to the invention were also able to achieve their intended purpose without showing acute toxicity against mammals such as mice and dogs as a result of the experiment.

In addition, in the case of clinically administering the compound of the present invention to obtain the desired anticoagulant effect and thrombolytic effect, the active compound (I) according to the present invention may be selected from at least one component selected from thrombolytic agents and platelet inhibitors. It may be administered simultaneously.

Thrombolytic agents that can be administered in combination with a compound of the present invention in this manner include t-PA, urokinase, Streptokinase, and the like, and platelet inhibitors include aspirin, ticlopidin, clopilo Gel (Clopidrogml), 7E3 monoclonal antibody, and the like.

However, preparations containing the compounds according to the invention for the treatment and prevention of blood clots are not limited to those described above, and any preparations useful for the compounds of the invention for the treatment and prevention of blood clots may be included.

The present invention is explained in more detail by the following Preparation Examples, Examples and Experimental Examples, but the present invention is not limited in any way by these.

[Production Example 1]

Synthesis of (S) -2- (N-t-butoxycarbonyl-amino) -pent-4-inoic acid

9.20 g (59.35 mmol) of N-acetyl-DL-2-amino-pent-4-inoic acid prepared according to the method described in the journal (Tetrahedron, 1991, 49, 5309) was dissolved in 300 ml of water, The pH was adjusted to 6.3 using aqueous sodium hydroxide solution. 0.2 g of acylase was added thereto and stirred at about 37 ° C. for 3 days. The reaction solution was filtered through Celite to remove the enzyme. The reaction solution was cooled to 0 ° C., adjusted to pH 1 with 6N hydrochloric acid, and washed several times with ethyl acetate. The aqueous solution was cooled back to 0 ° C., adjusted to neutral with 6N aqueous sodium hydroxide solution and the solution concentrated to 150 ml. The pH was adjusted to 10 again and 100 ml of 1,4-dioxane was added thereto. 8.4 g (38.53 mmol) of di-t-butyldicarbonate was added to the reaction solution, stirred at room temperature for 4 hours, and then distilled under reduced pressure to concentrate the solution and adjust the pH to 10. After washing twice with diethyl ether, the mixture was adjusted to pH 1 again, extracted with ethyl acetate, dried over anhydrous magnesium sulfate, and filtered. The filtrate was distilled off under reduced pressure to remove the solvent and dried under vacuum to afford 4.0 g (63% yield) of the title compound.

[Production Example 2]

Synthesis of (S) -2- (N-t-butoxycarbonyl-amino) -pent-4-inoic acid dimethylamide

2.0 g (9.4 mmol) of the compound obtained in Preparation Example 1 were dissolved in 60 ml of dimethylformamide (DMF), and 0.8 g (10.7 mmol) of dimethylamine hydrochloride and 1-hydroxybenzotriazole hydrate (HOBT) 1.4 g (10.4 mmol) was added and stirred until complete dissolution. After cooling the reaction solution to 0 ° C., 1.2 ml (10.0 mmol) of N-methylmorpholine and 2.0 g (10.4 mmol) of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDC) were added thereto. The reaction was slowly raised to room temperature and reacted for 4 hours. The solvent was distilled off under reduced pressure, and the residue was diluted with ethyl acetate, washed successively with saturated aqueous sodium hydrogen carbonate solution, diluted hydrochloric acid, and saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated and dried to give 1.9 g of a title compound (yield 83). %) Was obtained.

[Manufacture example 3]

Synthesis of (S) -2- (naphthalene-2-sulfonylamino) -pent-4-inoic acid dimethylamide

1.87 g (7.79 mmol) of the compound obtained in Preparation Example 2 were dissolved in 8 ml of dichloromethane, cooled to −10 ° C., and 5 ml of trifluoroacetic acid (TFA) was added thereto, stirred for 5 minutes, and the temperature was gradually decreased to room temperature. Raised. The reaction mixture was stirred for 30 minutes and distilled under reduced pressure to remove volatiles. The resulting residue was dried with a vacuum pump and 30 ml of DMF was added. After cooling to 0 ° C and adding 3 ml of N, N-diisopropylethylamine, the mixture was raised to room temperature and stirred for about 5 minutes. 1.77 g (7.81 mmol) of 2-naphthalenesulfonyl chloride was added thereto, stirred for 1 hour to complete the reaction, and distilled under reduced pressure to remove volatiles. The obtained residue was dissolved in ethyl acetate, washed twice with water, dried over anhydrous magnesium sulfate, filtered and concentrated. The residue was purified by column chromatography using ethyl acetate / hexane (2/1, volume ratio) as the eluent to obtain 1.70 g (yield 66%) of the title compound.

Example 1

Synthesis of (S) -2- (naphthalene-2-sulfonylamino) -5-phenyl-pent-4-inoic acid dimethylamide

100 mg (0.30 mmol) of the compound synthesized in Preparation Example 3 were added to a well-dried reaction vessel, and 21 mg (0.1 equiv) of bis (triphenylphosphine) pallidium (II) chloride and 6 mg (0.1 equiv) of copper iodide Was added. 34 µl (0.30 mmol) of iodinebenzene, 0.21 ml (5 equivalents) of triethylamine and 5 ml of acetonitrile were added thereto under nitrogen, followed by stirring at room temperature. After reacting for 1 hour, the solvent was removed by distillation under reduced pressure. The obtained residue was dissolved in a minimum amount of dichloromethane, and purified by column chromatography using ethyl acetate / hexane (2/1, volume ratio) as the eluent to obtain 70 mg (yield 57%) of the title compound.

Example 2

Synthesis of (S) -2- (naphthalene-2-sulfonylamino) -5-pyridin-4-yl-pent-4-inoic acid dimethylamide

100 mg (0.30 mmol) of the compound synthesized in Preparation Example 3 were added to a well-dried reaction vessel, 21 mg (0.1 equiv) of bis (triphenylphosphine) palladium (II) chloride and 6 mg (0.1 equiv) of copper iodide, And 64 mg (0.33 mmol) of 4-bromopyridine hydrochloride were added. 0.25 ml (6 equivalents) of triethylamine and 5 ml of acetonitrile were added thereto under nitrogen, followed by stirring while refluxing under heating. After reacting for 1 hour, the mixture was cooled to room temperature and distilled under reduced pressure to remove the solvent. The obtained residue was dissolved in a minimum amount of dichloromethane, and purified by column chromatography using ethyl acetate / hexane (2/1, volume ratio) as the eluent to obtain 60 mg (yield 49%) of the title compound.

Example 3

Synthesis of (S) -2- (naphthalene-2-sulfonylamino) -5-pyridin-3-yl-pent-4-inoic acid dimethylamide

66 mg (yield 54%) of the title compound was obtained in the same manner as in Example 2, except that 3-bromopyridine was used instead of 4-bromopyridine hydrochloride and 5 equivalents of triethylamine was used. It was.

Example 4

Synthesis of (S) -2- (naphthalene-2-sulfonylamino) -5-pyrimidin-5-yl-pent-4-inoic acid dimethylamide

Using 70-bromopyrimidine instead of 3-bromopyridine, 70 mg (yield 57%) of the title compound were obtained following the same method as in Example 3.

Example 5

Synthesis of 5- (6-amino-pyridin-3-yl)-(S) -2- (naphthalene-2-sulfonylamino) -pent-4-inoic acid dimethylamide

50 mg (yield 39%) of the title compound was obtained following the same method as Example 3 using 2-amino-5-bromopyrimidine instead of 3-bromopyridine.

Example 6

Synthesis of 5- (2-amino-pyridin-5-yl)-(S) -2- (naphthalene-2-sulfonylamino) -pent-4-inoic acid dimethylamide

52 mg (yield 41%) of the title compound was obtained following the same method as Example 3 using 2-amino-5-bromopyrimidine instead of 3-bromopyridine.

Example 7

Synthesis of (S) -2- (naphthalene-2-sulfonylamino) -5- (1H-pyrazol-4-yl) -pent-4-inoic acid dimethylamide

Using 4-bromopyrazole in place of 3-bromopyridine, 60 mg (yield 50%) of the title compound were obtained following the same method as in Example 3.

[Production Example 4]

Synthesis of (S) -2- (naphthalene-2-sulfonylamino) -5- (1-trityl-1H-imidazol-4-yl) -pent-4-inoic acid dimethylamide

The same as Example 3 using 4-iodo-1-trityl-imidazole prepared according to the method reported in the journal J. Org. Chem. 1981, 46, 1781 instead of 3-bromopyridine 120 mg (63% yield) of the title compound was obtained following the method.

Example 8

Synthesis of 5- (1H-imidazol-4-yl)-(S) -2- (naphthalene-2-sulfonylamino) -pent-4-inoic acid dimethylamide hydrochloride

120 mg (0.19 mmol) of the compound synthesized in Preparation Example 4 was dissolved in 3 ml of water and tetrahydrofuran (THF), respectively. Two drops of 6N hydrochloric acid was added thereto, and two drops of hydrochloric acid were added thereto while heating under reflux until the reaction was completed. After 30 minutes, the reaction mixture was cooled to room temperature and distilled under reduced pressure to remove THF. The residue was washed with ethyl acetate and the cloudy aqueous layer was filtered through celite. The filtrate was distilled off under reduced pressure to remove water. The remaining residue was purified by column chromatography using methanol / dichloromethane (1/19, volume ratio) as the eluent to give 70 mg (yield 85%) of the title compound.

Production Example 5

Synthesis of (S) -2- (naphthalene-2-sulfonylamino) -pent-4-inoic acid cyclopentyl-methyl-amide

(S) -2- (Nt-butoxycarbonyl-amino) -pent-4-inoic acid cyclopentyl- according to the same method as Preparation Example 2 using cyclopentyl-methylamine hydrochloride instead of dimethylamine hydrochloride Methyl-amide was synthesized. 5.0 g (17 mmol) of this compound were treated in the same manner as in Preparation Example 3, to obtain 4.4 g (yield 68%) of the title compound.

Example 9

Synthesis of (S) -2- (naphthalene-2-sulfonylamino) -5-phenyl-pent-4-inoic acid cyclopentyl-methyl-amide

43 mg (yield 36%) of the title compound was obtained in the same manner as in Example 1, using the compound synthesized in Preparation Example 5 instead of the compound synthesized in Preparation Example 3.

Example 10

Synthesis of (S) -2- (naphthalene-2-sulfonylamino) -5-pyridin-3-yl-pent-4-inoic acid cyclopentyl-methyl-amide

Instead of the compound synthesized in Preparation Example 3, using the compound synthesized in Preparation Example 5 to obtain 30 mg (25% yield) of the title compound purified in the same manner as in Example 3.

Example 11

Synthesis of 5- (6-amino-pyridin-3-yl)-(S) -2- (naphthalene-2-sulfonylamino) -pent-4-inoic acid cyclopentyl-methyl-amide

Using the compound synthesized in Preparation Example 5 instead of the compound synthesized in Preparation Example 3, 27 mg (yield 22%) of the title compound was obtained in the same manner as in Example 5.

[Manufacture example 6]

Synthesis of 3-iodo-N-t-butoxycarbonyl-aniline

0.55 mL (4.57 mmol) of 3-iodo-aniline was stirred with 3 mL of water and 6 mL of 1,4-dioxane. After cooling to 0 ° C., 5 ml of 1N aqueous sodium hydroxide solution was added, followed by 1.1 g (5.05 mmol) of di-t-butyldicarbonate. The reaction mixture was stirred at room temperature for 12 hours. Distillation under reduced pressure concentrated the solution, adjusted to pH 10, washed twice with diethyl ether and adjusted to pH 1 again. The reaction solution was extracted with ethyl acetate, dried over anhydrous magnesium sulfate and filtered.

The solvent was distilled off under reduced pressure and dried in vacuo to yield 0.82 g (56% yield) of the title compound.

[Manufacture example 7]

Synthesis of 5- [3- (N-t-butoxycarbonyl-amino) -phenyl]-(S) -2- (naphthalene-2-sulfonylamino) -pent-4-inoic acid cyclopentyl-methyl-amide

60 mg of the title compound purified according to the same method as Example 1 using the compound synthesized in Preparation Example 5 instead of the compound synthesized in Preparation Example 3, and the compound synthesized in Preparation Example 6 instead of iodobenzene. Yield 40%) was obtained.

Example 12

Synthesis of 5- (3-amino-phenyl)-(S) -2- (naphthalene-2-sulfonylamino) -pent-4-inoic acid cyclopentyl-methyl-amide hydrochloride

60 mg (0.10 mmol) of the compound synthesized in Preparation Example 7 was dissolved in 2.0 mL of 4N hydrochloric acid-dioxane, stirred at room temperature for 2 hours, and then distilled under reduced pressure to remove volatiles. The remaining residue was purified by column chromatography using methanol / dichloromethane (1/19, volume ratio) as the eluent to give 30 mg (59% yield) of the title compound.

Example 13

Synthesis of 5- (4-amino-phenyl)-(S) -2- (naphthalene-2-sulfonylamino) -pent-4-inoic acid cyclopentyl-methyl-amide trifluoroacetic acid salt

4-iodo-N-t-butoxycarbonyl-aniline was obtained according to the same method as Preparation Example 6 using 4-iodo-aniline instead of 3-iodo-aniline. This compound was treated in the same manner as in Production Example 7 to obtain the intermediate 5- [4- (Nt-butoxycarbonyl-amino) -phenyl]-(S) -2- (naphthalene-2-sulfonylamino) -pent 0.17 g (77% yield) of 4-ino-acid cyclopentyl-methyl-amide were obtained. 0.15 g (0.26 mmol) of this compound was dissolved in 1.5 mL of dichloromethane, cooled to −10 ° C., and 1.5 mL of trifluoroacetic acid (TFA) was added thereto. The reaction mixture was stirred for 30 minutes and distilled under reduced pressure to remove volatiles. The residue was purified by column chromatography using methanol / dichloromethane (1/99, volume ratio) as the eluent to afford 11 mg (7.2% yield) of the title compound.

Example 14

Synthesis of 5- (1H-imidazol-4-yl)-(S) -2- (naphthalene-2-sulfonylamino) -pent-4-inoic acid cyclopentyl-methyl-amide hydrochloride

Instead of the compound synthesized in Preparation Example 3, using the compound synthesized in Preparation Example 5 was carried out according to the same method as in Example 8 to obtain 22 mg (yield 52%) of the title compound.

[Manufacture example 8]

Synthesis of naphthalene-2-sulfonic acid [(S) -1- (piperidine-1-carbonyl) -but-3-ynyl] -amide

Intermediate (S) -2- (Nt-butoxycarbonyl-amino) -1-piperidin-1-yl-pent- according to the same method as Preparation Example 2 using piperidine instead of dimethylamine hydrochloride 4-yn-1-ones were synthesized. 0.96 g (3.43 mmol) of this compound were treated in the same manner as in Production Example 3 to obtain 0.78 g (61% yield) of the title compound.

Example 15

Synthesis of naphthalene-2-sulfonic acid [4- (3-amino-phenyl)-(S) -1- (piperidine-1-carbonyl) -but-3-ynyl] -amide hydrochloride

Using the compound of Preparation Example 8 in place of the compound of Preparation Example 5, the process was carried out according to the same method as in Example 12, to obtain 55 mg (yield 96%) of the title compound.

Example 16

Synthesis of naphthalene-2-sulfonic acid [4- (1H-imidazol-4-yl)-(S) -1- (piperidine-1-carbonyl) -but-3-ynyl] -amide hydrochloride

Using the compound synthesized in Preparation Example 8 instead of the compound synthesized in Preparation Example 3 was carried out according to the same method as in Example 8 to obtain 16 mg (yield 46%) of the title compound.

Example 17

Synthesis of (S) -2- (naphthalene-2-sulfonylamino) -hex-4-inoic acid cyclopentyl-methyl-amide

50 mg (0.13 mmol) of the compound synthesized in Preparation Example 5 was dissolved in 3 ml of dried THF and cooled to -78 ° C. 0.3 ml (0.3 mmol) of 1.0M solution of lithium hexamethyldisilazide (LHMDS) dissolved in hexane was added thereto, the temperature was gradually raised to 0 ° C, stirred for 5 minutes, and then cooled to -78 ° C. Into another well-dried container, 10 µl (0.16 mmol) of methyl iodide and 0.5 ml of dry THF were added. The resulting solution was added to the first reaction vessel and stirred. After 1 hour, water was added to the reaction mixture and the reaction mixture was raised to room temperature to complete the reaction. The reaction solution was distilled off under reduced pressure, and extracted with water and ethyl acetate. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated. The residue was purified by column chromatography using ethyl acetate / hexane (1/1, volume ratio) as the eluent to obtain 48 mg (yield 92%) of the title compound.

[Manufacture example 9]

7- (1,3-dioxo-1,3-dihydro-isoindol-2-yl)-(S) -2- (naphthalene-2-sulfonylamino) -hept-4-inoic acid cyclopentyl- Synthesis of Methyl-amide

The procedure was carried out in the same manner as in Example 17 using N- (2-bromoethyl) -phthalimide instead of methyliodide to give 77 mg (yield 30%) of the title compound.

Example 18

Synthesis of 7-amino- (S) -2- (naphthalene-2-sulfonylamino) -hept-4-inoic acid cyclopentyl-methyl-amide

77 mg (0.14 mmol) of the compound synthesized in Preparation Example 9 were dissolved in 1 ml of ethanol. 20 µl (0.33 mmol) of 80% hydrazine hydrate was added thereto, and the mixture was stirred while refluxing under heating. The same amount of hydrazine hydrate was added at 2 hour intervals until the reaction was complete, yielding a white solid. Stirred for 6 hours, cooled to room temperature and filtered. The filtrate was distilled off under reduced pressure to remove the solvent, and the residue was purified by column chromatography using methanol / dichloromethane (1/33, volume ratio) as the eluent to obtain 30 mg (yield 50%) of the title compound.

Experimental Example

Inhibitory Activity of Thrombin Inhibitor

The enzymatic inhibitory effect of the thrombin inhibitor, a compound of formula (I) according to the present invention, was determined by the rate constant Ki measured by spectrophotometry, using a substrate that reacted with an enzyme to produce color. The rate constant Ki represents the extent to which the inhibitor inhibits the enzyme activity when the enzyme, substrate and inhibitor are added simultaneously. Hydrolysis of thrombin-based substrate, Chromozym TH (Tosyl-Gly-Pro-Arg-4-nitroanilide acetate), produces para-nitroaniline. The amount of yellow para-nitroaniline thus produced was measured as a change in absorbance over time. From this rate, the activity of the enzyme can be measured, and the inhibitory ability of the inhibitor can be measured.

The inhibitory ability of the inhibitor against thrombin was determined by the following method.

After adding 1160 μl of 0.1M Tris buffer solution (pH 7.8) containing 150 mM NaCl, 0.1% PEG 8000 (polyethylene glycol, molecular weight about 8,000) to a 1.5 ml cuvette, it was added to a chromatin substrate, Chromozyme TH (Chromozym TH). : 225 [mu] l of Tosyl-Gly-Pro-Arg-4-nitroanilide acetate) 0.1 mM solution was added. Chromozyme TH was dissolved in dimethylsulfoxide (DMSO) so as to have a concentration of 10 mM, and diluted with the buffer solution at a concentration of 0.1 mM. The thrombin inhibitor of the present invention was dissolved in dimethyl sulfoxide to a concentration of 10 mg / ml, and then diluted with Tris buffer solution having the composition mentioned above to 0.1 mg / ml, 0.01 mg / ml, 0.001 mg / ml. After the preparation was taken so that the amount of inhibitor was in the range of 0 to 10 µg, the Tris buffer solution was added to the final volume of 100 µl and added to each cuvette.

At room temperature, 15 µl of a sothrombin solution dissolved in the Tris buffer solution at a concentration of 0.1 mg / ml was added to the cuvette containing the reaction solution, and the enzyme hydrolysis reaction was started. The amount of para-nitroaniline produced by the reaction for 2 minutes from the moment of addition of the enzyme was monitored by a change in absorbance at 381 nm to draw a continuous spectrum of reaction time versus absorbance. The experiment was repeated at various inhibitor concentrations to obtain continuous spectra.

After obtaining the initial velocity Vi from the slope within the initial 30 seconds of the reaction time in each spectrum, a graph of the inverse of the initial velocity versus the inhibitor concentration (1 / Vi) is shown.

After calculating the first equation that satisfies the points on the graph, Ki was calculated from the x segment of the equation using the enzyme reaction equation. The K _M value used in this calculation was 8.3 μM, which was obtained by varying the substrate concentration at a constant enzyme concentration.

Table 1 shows the enzyme activity inhibitory capacity of each inhibitor measured for thrombin as Ki value.

Claims (10)

Compounds represented by the following general formula (I) and pharmaceutically acceptable salts, hydrates, solvates and isomers thereof: Wherein R ¹ is in each occurrence a substituted or unsubstituted aryl or heteroaryl, R ² and R ³ may be the same or different from each other, and each independently hydrogen, lower alkyl, cycloalkyl, or in each case Aryl or heteroaryl substituted or unsubstituted, R ² and R ³ are bonded together with the nitrogen atom to which they are attached to be substituted or unsubstituted by lower alkyl, carboxyl or carboalkoxy, and Containing 4 to 8-membered rings, R ⁴ represents lower alkyl, lower aminoalkyl, or in each case substituted or unsubstituted aryl, arylalkyl, heteroaryl or heteroarylalkyl. The compound of claim 1, wherein R ¹ represents aryl and R ² and R ³ each independently represent hydrogen, lower alkyl or cycloalkyl, or R ² and R ³ together with the nitrogen atom to which they are attached a 6-membered ring Wherein R ⁴ represents lower alkyl or aryl or heteroaryl unsubstituted or substituted by an amino group. 3. A compound according to claim 2, wherein R ¹ represents naphthyl and R ² and R ³ each independently represent hydrogen, methyl or cyclopentyl, or R ² and R ³ are bonded together with the nitrogen atom to which they are attached Forming a ferridine group, wherein R 4 represents phenyl, pyridyl, pyrimidinyl, pyrazolyl or imidazolyl unsubstituted or substituted by a methyl or amino group. (S) -2- (naphthalene-2-sulfonylamino) -5-phenyl-pent-4-inoic acid dimethylamide according to claim 3, (S) -2- (naphthalene-2-sulfonylamino) -5-pyridine-4-pent-4-inoic acid dimethylamide, (S) -2- (naphthalene-2-sulfonylamino) -5-pyridin-3-yl-pent-4-inoic acid dimethylamide, ( S) -2- (naphthalene-2-sulfonylamino) -5-pyrimidin-5-yl-pent-4-inoic acid dimethylamide, 5- (6-amino-pyridin-3-yl)-(S) -2- (naphthalene-2-sulfonylamino) -pent-4-inoic acid dimethylamide, 5- (2-amino-pyridin-5-yl)-(S) -2- (naphthalene-2-sulfonylamino ) -Pent-4-inoic acid dimethylamide, (S) -2- (naphthalene-2-sulfonylamino) -5- (1H-pyrazol-4-yl) -pent-4-inoic acid dimethylamide, 5 -(1H-imidazol-4-yl)-(S) -2- (naphthalene-2-sulfonylamino) -pent-4-inoic acid dimethylamide hydrochloride, (S) -2- (naphthalene-2-sul Ponylamino) -5-phenyl-pent-4-inoic acid cyclopentyl-methyl-amide, (S) -2- (naphthalene-2-sulfonylami ) -5-pyridin-3-yl-pent-4-inoic acid cyclopentyl-methyl-amide, 5- (6-amino-pyridin-3-yl)-(S) -2- (naphthalene-2-sulfonyl Amino) -pent-4-inoic acid cyclopentyl-methyl-amide, 5- (3-amino-phenyl)-(S) -2- (naphthalene-2-sulfonylamino) -pent-4-inoic acid cyclopentyl -Methyl-amide hydrochloride, 5- (4-amino-phenyl)-(S) -2- (naphthalene-2-sulfonylamino) -pent-4-inoic acid cyclopentyl-methyl-amide trifluoroacetic acid salt, 5- (1H-imidazol-4-yl)-(S) -2- (naphthalene-2-sulfonylamino) -pent-4-inoic acid cyclopentyl-methyl-amide hydrochloride, naphthalene-2-sulfonic acid [4 -(3-Amino-phenyl)-(S) -1- (piperidine-1-carbonyl) -but-3-ynyl] -amide hydrochloride, naphthalene-2-sulfonic acid [4- (1H-imidazole- 4-yl)-(S) -1- (piperidine-1-carbonyl) -but-3-ynyl] -amide hydrochloride, (S) -2- (naphthalene-2-sulfonylamino) -hex- 4-ino-acid cyclopentyl-methyl-amide, and 7-amino- (S) -2- ( Program talren-2-sulfonyl amino) hept-4-Ino acid cyclopentyl-compound is selected from the group consisting of polyamide-methyl. A process for preparing a compound of formula (I) and salts thereof, characterized by reacting an acetylene compound of formula (II) with a compound of formula (III): Wherein R ¹ is in each occurrence a substituted or unsubstituted aryl or heteroaryl, R ² and R ³ may be the same or different from each other, and each independently hydrogen, lower alkyl, cycloalkyl, or in each case Aryl or heteroaryl substituted or unsubstituted, R ² and R ³ are bonded together with the nitrogen atom to which they are attached to be substituted or unsubstituted by lower alkyl, carboxyl or carboalkoxy, and Containing 4 to 8-membered rings, R ⁴ represents lower alkyl, lower aminoalkyl, or in each case substituted or unsubstituted aryl, arylalkyl, heteroaryl or heteroarylalkyl, and X is Halogen is represented. The acetylene compound of formula (II) according to claim 5, wherein R ⁴ is substituted or unsubstituted aryl, arylalkyl, heteroaryl or heteroarylalkyl in the presence of copper iodide and palladium catalyst. A method characterized by reacting with a compound. A process according to claim 5, wherein the acetylene compound of formula (II) is treated with a strong base and then reacted with a compound of formula (III) wherein R ⁴ represents lower alkyl or lower aminoalkyl under cooling. The method according to claim 5, wherein the compound of formula (II) is a compound prepared by coupling a compound of formula (4) with a compound of formula (5). Wherein R ¹ , R ² and R ³ are as defined in claim 4. Compounds of the following general formula (II): Wherein R ¹ , R ² and R ³ are as defined in claim 1. A composition for preventing coagulation and treating thrombosis, comprising a compound of formula (I) according to any one of claims 1 to 4 as an active ingredient together with a pharmaceutically acceptable carrier.